Preferential exclusion of sucrose from recombinant interleukin-1 receptor antagonist: role in restricted conformational mobility and compaction of native state

Understanding the mechanism for sucrose-induced protein stabilization is important in many diverse fields, ranging from biochemistry and environmental physiology to pharmaceutical science. Timasheff and Lee [Lee, J. C. & Timasheff, S. N. (1981) J. Biol. Chem. 256, 7193-7201] have established that thermodynamic stabilization of proteins by sucrose is due to preferential exclusion of the sugar from the protein's surface, which increases protein chemical potential. The current study measures the preferential exclusion of 1 M sucrose from a protein drug, recombinant interleukin 1 receptor antagonist (rhIL-1ra). It is proposed that the degree of preferential exclusion and increase in chemical potential are directly proportional to the protein surface area and that, hence, the system will favor the protein state with the smallest surface area. This mechanism explains the observed sucrose-induced restriction of rhIL-1ra conformational fluctuations, which were studied by hydrogen-deuterium exchange and cysteine reactivity measurements. Furthermore, infrared spectroscopy of rhlL-1ra suggested that a more ordered native conformation is induced by sucrose. Electron paramagnetic resonance spectroscopy demonstrated that in the presence of sucrose, spin-labeled cysteine 116 becomes more buried in the protein's interior and that the hydrodynamic diameter of the protein is reduced. The preferential exclusion of sucrose from the protein and the resulting shift in the equilibrium between protein states toward the most compact conformation account for sucrose-induced effects on rhIL-1ra.     

Airway stents. Present and future

Tracheobronchial stent insertion is a relatively new technique used to palliate or cure central airways obstruction. When performed by experienced thoracic endoscopists, this procedure is both safe and effective, even if complications of indwelling stents may require repeat endoscopic intervention. Although proven clinically beneficial to many patients, airway prostheses have not yet been the subject of large comparative case studies or randomized controlled investigations. Reports are often experiential and anecdotal. Further research is needed to determine the effects of stenting on survival in patients with malignant airway disease and its degree of success in patients with benign airway strictures. Only then will airway stents truly deserve their place alongside other therapies for tracheobronchial obstruction.     

Microwave signal generation using an optical self-heterodyne technique

A new technique for mixing optical waves to generate microwave-frequency signals using an injection laser diode and a fibre-optic interferometer is reported. A beat signal is produced by interfering light from the laser with light from the same laser emitted earlier at a different frequency. The interferometer consists of an evanescent-field fibre coupler joined to a length of single-mode optical fibre. The laser is tuned by pulsed or bipolar current waveforms superimposed on a DC bias current. Feedback from an external cavity stabilises the laser frequency and reduces its linewidth. Efficient mixing at difference frequencies from 250 MHz to 2 GHz is obtained, with spectral widths of less than 1% of the centre frequency.     

Artefactual cleavage of E coli H-NS by OmpT

In the bacterium Escherichia coli, H-NS-(H1, H1a) is a heat-stable protein with a molecular mass of 15.5 kDa involved in nucleoid organisation and gene regulation linked to certain signal transduction pathways. We have shown that, following addition of preparations of everted inner membrane vesicles, heat-stable cleavage products of approximately 10 kDa of H-NS are formed in vitro from newly synthesised, radio-labelled H-NS and from purified H-NS. The 15.5 kDa protein and its cleavage products were also recovered from a minicell system. These results raised the possibility that cleavage of H-NS is physiologically significant. However, the cleavage of H-NS observed appears to occur during cell breakage and to depend on the method of protein extraction and the presence of the outer membrane protease, OmpT. Nevertheless, the results indicate that H-NS may contain at least two separate domains with cleavage occurring between these domains at a preferred OmpT site. Failure to take account of H-NS cleavage in sample preparation and analysis can lead to serious underestimation of H-NS levels.     

Paroxysmal atrial fibrillation

Paroxysmal atrial fibrillation is the most frequent of cardiac arrhythmias and its frequency increases with age. Its diagnosis is difficult since the symptoms are often absent or misleading. It is based on electrocardiogram performed during the attack, and this has been made easier by the Holter technique or by sequential recordings triggered by the patient himself. It is only in special cases, when percritical recordings could not be made that intracavitary electrophysiological study is used to obtain indirect arguments in favour of the diagnosis. All heart diseases can be attended by this cardiac rhythm disorder which usually marks an unfavourable turn in the course of the disease, with an increased risk of sudden death in the end. But in most cases the only disorder found is a common arterial hypertension. Atrial fibrillation carries a risk of peripheral, and in particular cerebral, embolism evaluated at 5% per annum. This risk is high in patients with cardiopathy, but it also exists even with a normal heart. The value of anticoagulants has been demonstrated by large controlled studies, but it is important to know that a very strong anticoagulant treatment is unnecessary to reduce the risk of embolism. Only antiarrhythmic agents can maintain the patients in sinus rhythm for as long as possible before the passage to permanent fibrillation. Although no beneficial effect on mortality has been proved, these drugs are indicated for patients who have very marked symptoms or a major cardiopathy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Improved accuracy of burn wound assessment using laser Doppler

The utility of the laser Doppler for determining burn depth has been questioned because of problems with technology and methodology. This study prospectively evaluates the ability of a new laser Doppler technique to predict burn healing time. Using the Periflux System 4000 laser Doppler, readings were taken on 305 burns (147 patients) on postburn day 3 or 4. Sixty-six wounds were used to derive a predictive function (phase I) and 152 wounds were used to test the function (phase II). Blood flow dynamics (flux), microvascular dilation capacity of the wounds to beat stress, and flow motion wave pattern (vasomotion) were studied using the laser Doppler, and seven parameters were evaluated to determine their relative contribution to the prediction of healing time. These parameters are hyperemic flux (flux value after heating to 42 degrees C), average hyperemic wave amplitude (AHWA), number of average flux units >100(F100), number readings with wave amplitude 75 (A5), average flux change (AFC), percentage of average flux increase, and relative flow capacity (RFC = AFC/average hyperemic flux). After readings were made, the wounds were observed and divided into two groups: those that healed in less than 14 days and those that healed or were grafted after 14 days. A step-wise discriminant analysis was used to assess the relative contribution of the Doppler-derived measures to healing time prediction. AHWA, F100, and RFC were included in the final discriminant function explaining 72% of the healing time variance (Wilks' lambda value 0.28; p value <0.0001). Predicted outcome = 0.05(AHWA) + 0.31(F100) + 5.0(RFC) - 2.3. With this derived function, there is 94% accuracy in the prediction of burn wound healing time compared with a physician predictive accuracy of 70%.     

Age-related changes in cognitive ERPs of attenuation

The purpose of this article is to deliberate the moral and legal dilemma entailed in the weapon of the labour strike as a pressure tactic on the Israeli Finance Ministry regarding job slots, budgets and, in effect, violating the collective agreement signed by the nurses and impairing patients' treatment, as opposed to refraining from striking and suffering the heavy burden of work, the lack of trained personnel, low wages, and the inability to give patients proper, high quality treatment.